CN100340288C - Inhibitors for continuous activation of calcineurin - Google Patents
Inhibitors for continuous activation of calcineurin Download PDFInfo
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- CN100340288C CN100340288C CNB2003801005939A CN200380100593A CN100340288C CN 100340288 C CN100340288 C CN 100340288C CN B2003801005939 A CNB2003801005939 A CN B2003801005939A CN 200380100593 A CN200380100593 A CN 200380100593A CN 100340288 C CN100340288 C CN 100340288C
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Abstract
The object of the present invention is to provide a suppressant for neuronal cell death effective for various diseases which inhibits a constitutive active forming of calcineurin and has less side effect. An inhibitor of constitutive active forming of calcineurin, more specifically a drug which inhibits cleavage of calcineurin subunit A (CaNA) by calpain. Examples thereof include peptides having the amino acid sequence of FDGATAAARKEVIRNK (SEQ No. 1) and REESESVLTLKGLTPTG (SEQ No. 2).
Description
Technical field
The present invention relates to the persistent activation inhibitor of calcineurin.Say in detail the present invention relates to and suppress the medicine that calcineurin A subunit (CaNA) is cut off by Calpain.
Background technology
Calcineurin (calcineurin) is the dephosphorylase that is activated that depends on calcium and calmodulin, CaM, is by calcineurin A subunit (CaNA) with active center and the complex that constitutes as the calcineurin B subunit (CaNB) of regulatory factor.Calcineurin is owing to suppress combining of active center and substrate by self suppressing domain, so be non-active form usually in cell.If intracellular calcium concentration rises, the structural change of calcineurin self suppresses domain and opens (active form), and substrate just can combine with the active center.In addition, if calcium concentration reduces, calcineurin returns to nonactive type again.As mentioned above, calcineurin is depended on calcium concentration by reversible activation as can be known.
Reported that calcineurin had important effect (with reference to Asai Akio etc., J.Biological Chemistry, the 274th volume, P.34450,1999) to nerve cell death in 1999.The special inhibitor (immunosuppressant FK506 and cyclosporin A) of also having reported calcineurin in addition also suppress nerve cell death (with reference to Morioka Motohiro etc., Progressin Neurobiology, the 58th volume, P.1,1999 and Springer Joe E etc., TheJournal of Neuroscience, the 20th volume, P.7246,2000).According to a lot of reports that comprise these documents; about the nerve cell death of in cerebral ischemia and spinal cord injury etc., being seen; proposed now because the excitement of neurocyte; N-methyl-D-aspartate receptor (nmda receptor) abnormal activation as one of glutamate receptor; a large amount of calcium causes calcineurin to be activated in the calcium receptor flows into cell, brings out the mechanism of cell death.Yet the inflow of this calcium is instantaneous, and the state of activation of calcineurin can not last very long.Therefore, imagined that existing other is not one to cross calcium concentration in the sexual cell and rise the calcineurin that causes by long-term activatory mechanism, but relevant such mechanism is not reported also.
As mentioned above, for the inhibition of nerve cell death, specifically for the inhibition of ischemic and excitatory neuron cell death, known immunosuppressant FK506 and cyclosporin A are effective.Yet known these medicines are owing to suppressing all information transmission that calcineurin participates in, so side effect big (for example nephrotoxicity and diabetes etc.).Therefore look forward to suppressing long calcineurin activation strongly and compare the inhibitor of the littler nerve cell death of side effect with medicine in the past.
Summary of the invention
The object of the invention is to solve above-mentioned such problem, provide side effect little, to the effective nerve cell death inhibitor of various disease.
In order to solve above-mentioned problem, constantly study, the result has obtained following opinion.
1) by giving the glutamate induction nerve cell death, confirms the fracture of calcineurin A subunit (CaNA).
2) between the 392nd (arginine) and 393 (lysine) of the place of incision of CaNA in aminoacid sequence and between the 421st and 425.
3), show that calcineurin has calcium and calmodulin, CaM dependent/non-dependent activity if calcineurin is cut off at above-mentioned position.
4) above-mentioned fracture be by think the information transmission that belongs to different with calcineurin by way of Calpain cause, can be suppressed by the Calpain inhibitor.
5) if will contain the peptide of 392～393 amino acid residues of CaNA or the peptide that contains 421～425 amino acid residues of CaNA imports in the neurocyte, will suppress to give the nerve cell death that glutamic acid causes.
According to above opinion, exploitation is the calcineurin activation that causes but with the cell death inhibitor of the activatory mechanism of secular calcineurin as target, finished the present invention because instantaneous intracellular calcium concentration rises not to be.Promptly the present invention relates to:
(1) inhibitor that cut off by Calpain of the inhibition CaNA of the peptide of a kind of peptide that contains sequence 1, sequence 2 and/or their analog,
(2) more than one cut-out inhibitor of stating (1) described CaNA are the nerve cell death inhibitor of effective ingredient,
(3) more than one cut-out inhibitor of stating (1) described CaNA inhibitor that is the dementia progression of disease of effective ingredient, and
(4) a kind of cell of the cut-out inhibitor that contains above-mentioned (1) described CaNA and the additive of brain section culture medium.
The simple declaration of accompanying drawing
The photo of the inhibition effect of the inhibition CaNA cut-out that the cut-out of the CaNA that Fig. 1 is expression when adding the glutamate induction nerve cell death and the inhibitor by CaNA of the present invention produce.The glutamic acid result of the sample of incubation after 3 hours is afterwards added in Fig. 1 (a) expression, the result of the sample of incubation after 24 hours after Fig. 1 (b) expression interpolation glutamic acid.At Fig. 1 (a) with (b), with 1 expression contrast, with 2 expressions only add glutamic acid sample, only add the sample that cuts off peptide for inhibiting, add glutamic acid and cut off the samples of peptide for inhibiting with 3 expressions with 4 expressions.
Fig. 2 represents owing to add the number of the neurocyte of glutamate induction nerve cell death.
The specific embodiment
Inhibition calcineurin A subunit (CaNA: the sequence 3) inhibitor that is cut off by Calpain of the present invention, be the peptide that contains sequence 1, the peptide of sequence 2 and/or the chemical compound of their analog, refer to and suppress between 392～393 amino acid residues that Calpain cuts off CaNA and position between 421～425 amino acid residues, suppress the lasting activatory material of calcineurin.
Analog as the peptide of above-mentioned sequence 1, for example lacked, replaced and/or inserted other aminoacid sequences by the part in the aminoacid sequence of sequence 1 peptide, or in the terminal aminoacid sequence formation of adding other aminoacid sequence of peptide, and comprise the peptide that suppresses Calpain cut-out CaNA.For example comprise in the analog of the peptide of sequence 1 of the present invention that the amino acid residue Lys that the 9th amino acids residue A rg in the peptide sequence of sequence 1 is replaced into peptide behind the Lys and/or the 10th is replaced into the peptide of Arg.
Analog as the peptide of above-mentioned sequence 2, for example lacked, replaced and/or inserted other aminoacid sequences by the part in the aminoacid sequence of sequence 2 peptides, or in the terminal aminoacid sequence formation of adding other aminoacid sequence of peptide, and comprise the peptide that suppresses Calpain cut-out CaNA.In the analog of the peptide of sequence 2 of the present invention, for example comprise that the 11st amino acids residue Lys in the peptide sequence of sequence 2 is replaced into the peptide behind the Arg.
The inhibition activity that the inhibition CaNA of the peptide analogues of sequence 1 or sequence 2 (similar peptide) cuts off can resemble to be estimated following.That is, (contain 0.1 μ M or the similar peptide of 10 μ M, 5 μ M purification Calpains, 20mM Tris-HCl (pH7.4) and 1mMCaCl in preparation testing liquid
2) after, add purification calcineurin (final concentration 1 μ M), under 30 ℃, carried out incubation 1 hour.By 10%SDS-PAGE reactant liquor is carried out electrophoresis on gel then, gel dyes with the bright basket of coomassie.Undertaken quantitatively can estimating by molecular weight 45kDa that coloration result is seen and the Calpain dependency fragmentation calcineurin of 48kDa.
Above-mentioned peptide can use well-known methods such as the solid phase of Boc method or Fmoc method or liquid phase be synthetic to make.In addition, the peptide of making so also can precipitate with ether-well-known methods such as Filtration, high performance liquid chromatography (HPLC), Perfusion Chromatography carry out purification.
Inhibition Calpain of the present invention cuts off the inhibitor of CaNA so long as contain the peptide of sequence 1, the peptide of sequence 2 and/or their analog, also can contain other chemical compound.Other chemical compound, for example the protein guiding structure territory (PTD that constitutes of 11 amino acid residues that contain in poly arginine peptide (for example, the poly arginine peptide that constitutes by 5 arginine), the TAT protein by HIV virus; Lead-in signal peptide in the cell of sequence 4) such 7～30 formations that contain arginine more than 50% or lysine, or as the straight linear polyethylene imines (PEI) of cationic water-soluble polymer etc.These chemical compounds utilize (i) by the synthetic synthetic method that is connected to the peptide and/or their analog of sequence 1 or sequence 2 of common peptide, or (ii) make one of them peptide connect the divalent cross-linking agent, the end of another peptide connects cysteine residues, method by making two reactive polypeptides etc. can combine (fusion) with the peptide and/or their analog of sequence 1 or sequence 2.The cut-out inhibitor of the CaNA that obtains like this can use above-mentioned purification process to carry out purification.
Nerve cell death inhibitor of the present invention contains the inhibitor that above-mentioned inhibition Calpain cuts off CaNA as effective ingredient, refers to suppress the long inductive medicine of nerve cell death in the neurocyte.Nerve cell death inhibitor also can be used as prevention or the therapeutic agent use with the nerve cell death diseases associated.Nerve cell death inhibitor of the present invention preferably contains the cut-out inhibitor of the CaNA that is made of lead-in signal peptide in the peptide of sequence 2 and the cell, more preferably contains the cut-out inhibitor of the CaNA that is made of lead-in signal peptide in the peptide of the peptide of sequence 1, sequence 2 and the cell.Refer to the prevention of nerve cell death diseases associated or therapeutic agent and to contain the medicine that is useful on the nerve cell death inhibitor of the effective dose of the prevention of nerve cell death diseases associated or treatment.As with the nerve cell death diseases associated, as intracerebral hemorrhage, spinal cord injury, parkinson disease and epilepsies etc. such as Alzheimer's disease, dementia disease, cerebral ischemia diseases, subarachnoid hemorrhages.
As the medicine-feeding way of nerve cell death inhibitor of the present invention, can enumerate as oral administration, directly administration etc. in intravenously administrable and brain, consider that from burden, side effect viewpoint oral administration is better to the patient.
The dosage form of nerve cell death inhibitor of the present invention can suitably be set according to medication.Specifically, as liquors such as aqueous solution, Emulsion, suspension, tablet and capsule etc.For example, during oral administration, preferred tablet or capsule, directly preferred liquid agent during administration in intravenously administrable or brain.In the preparationization of nerve cell death inhibitor of the present invention, the various additives that can use common those skilled in the art of being complementary with this dosage form all to use.For example prevent that oxidant, pH from adjusting agent, antiseptic etc.
The dosage of above-mentioned nerve cell death inhibitor can suitably be set according to medication, suitable patient's age, body weight and condition of illness etc.For example, be scaled the inhibitor that inhibition Calpain of the present invention cuts off CaNA, preferably more than 0.1mg/kg every day, more preferably more than the 1mg/kg.Dosage exists CaNA and cuts off the tendency that the inhibition effect reduces by half than 0.1mg/kg after a little while.In addition, be scaled the inhibitor that inhibition Calpain of the present invention cuts off CaNA, preferably below 100mg/kg every day, more preferably below the 20mg/kg.When dosage surpasses 100mg/kg, exist to show Cytotoxic tendency.The administration of nerve cell death inhibitor of the present invention can be carried out by one or many.
The additive of cell culture medium of the present invention or brain section culture medium refers to the additive of the culture medium of the cut-out inhibitor that contains CaNA at least.The additive of cell culture medium of the present invention preferably contains the cut-out inhibitor of the CaNA that is made of lead-in signal peptide in the peptide of sequence 2 and the cell, more preferably contains the cut-out inhibitor of the CaNA that is made of lead-in signal peptide in the peptide of the peptide of sequence 1, sequence 2 and the cell.
When being applied to cultured cell, cut off the addition of the cut-out inhibitor of CaNA as inhibition Calpain of the present invention, at cell concentration 1 * 10
5Preferred 0.01～the 100nmol/ml of the culture fluid of cell/ml, more preferably 0.1～10nmol/ml.When addition was less than 0.01nmol/ml, the effect of the cut-out inhibitor of CaNA had the tendency that reduces by half, and surpassed the 100nmol/ml existence and showed Cytotoxic tendency.
Below according to embodiment the present invention is described in more detail, but the present invention is not subjected to the restriction of these embodiment.
Cut-out inhibitor as CaNA of the present invention, for FDGATAAARKEVIRNK (sequence 1) and REESESVLTLKGLTPTG (sequence 2) are imported in the cultured cell, make and as followsly added the oligopeptide of lead-in signal peptide (10 arginine) (production of peptide institute company) in the cell at N-terminal.
RRRRRRRRRRFDGATAAARKEVIRNK (sequence 5)
RRRRRRRRRRREESESVLTLKGLTPTG (sequence 6)
(preparation of neurocyte)
Behind young the 18th day cerebral hippocampal of Wister rat tire extraction, handled 15 minutes in 37 ℃ with containing 0.05% tryptic PBS.After using glass pipet that neurocyte is disperseed, cultivating 1 * 10 in the diameter 3.5cm culture dish with poly--D-lysine bag quilt in advance
6Individual cell.Culture medium is used and has been added B27supplement (0.03ml; Invitrogen, Inc. company produces), penicillin (final concentration 100 units/ml; Invitrogen company produces) and streptomycin (final concentration 100 μ g/ml; Invitrogen company produces) 3ml Neuro Basa1 culture medium (production of Invitrogen company), in CO2 gas incubator (5%CO2,37 ℃), cultivate.
(interpolation of peptide)
Cultivating beginning after 10 days, adding final concentration in culture fluid is the above-mentioned sequence 5 of 1 μ M and 6 peptide, in CO2 gas incubator (5%CO
2, 37 ℃) in cultivate.Add after 3 hours, add the glutamic acid of final concentration 500 μ M, cultivated 15 minutes.Exchange culture fluid then, cultivate again.After 3 hours and 24 hours, reclaim cell after adding glutamic acid, pair cell carries out Ultrasonic Pulverization in 1%SDS solution, adds the SDS-PAGE buffer.Same sample is behind the SDS-PAGE gel electrophoresis, and (rabbit anteserum, Santa CruzBiotechnology, Inc company produces) carries out immunoblotting (Western blotting) with the antibody of discerning CaNA.In addition, in the present embodiment, use the cell sample that does not add glutamic acid and cut off peptide for inhibiting in contrast.As parallel, also make the cell sample that only adds glutamic acid and only add the cell sample that cuts off peptide for inhibiting in addition.
The result as shown in Figure 1.In adding the glutamic acid group, confirm the fragmentation of calcineurin.And in importing the neurocyte that cuts off peptide for inhibiting, owing to add glutamic acid, the cut-out of common generable calcineurin is suppressed.
Similarly to Example 1, neurocyte is cultivated, it is 1 μ M that the peptide that adds sequence 5 as described above and 6 then similarly to Example 1 in the neurad cell makes final concentration.In contrast, add calcineurin inhibitors FK506 (Off ジ サ ワ Pharmaceutical Co., Ltd; Final concentration 1 μ M) or Calpain inhibitor ALLM (Merk company produces; Final concentration 25 μ M).Each medicine incubation after 2 hours, is added the glutamic acid of final concentration 500 μ M, incubation 15 minutes.Then, the exchange culture fluid is cultivated again.After adding back 3 hours, 6 hours, 12 hours or 24 hours, each neurocyte is fixed with 4% paraformaldehyde.Then, in order to identify the neurocyte that cell death takes place, carry out TUNEL dyeing (Roche Diagnostics, Inc produces).The TUNEL positive cell is counted.
The result as shown in Figure 2.In giving the glutamic acid group, along with passage of time after the administration, the neurocyte number that cell death takes place rises.Distinguish that cutting off peptide for inhibiting has the effect of inhibition because of the nerve cell death of glutamate induction.In addition, its effect has the effect with FK506, ALLM equal extent.
Reference example 1
To from Medulla Bovis seu Bubali, in 1 μ M calmodulin, CaM (Merck, Inc produces) and/or 1 μ M m-Calpain (Merck, Inc produces) reactant liquor, (contain 20mM Tris-HCl, pH7.4,1mM CaCl by the calcineurin 1 μ M of purification
2, 1mM MgCl
2) reacted respectively 1 hour in 30 ℃, carry out the 12%SDS-PAGE gel electrophoresis then after, with the bright basket of coomassie this gel is dyeed.
When the result does not add Calpain, see the size of purification CaNA 60Kda on running gel.This is big or small consistent with the CaNA's that up to the present reports.And when adding calmodulin, CaM and Calpain, do not see band at 60Kda, 48 and 45Kda seen band.If, confirm to be cut into the CaNA of 45Kda size only with the Calpain reaction.
The above results shows that calcineurin has been cut off by Calpain.
The result who determines the place of incision of Calpain cut-out CaNA in addition shows, 45Kda cuts off albumen and is made of the aminoacid before the 392nd of aminoacid sequence, the cut-out albumen of 48Kda by before 421, before 422, before 423 and 424 before aminoacid constitute.
The invention provides the cut-out inhibitor of CaNA.CaNA of the present invention cuts off inhibitor owing to can suppress the irreversible activation of calcineurin, and all can suppress the nerve cell death that brought out by this activation.In addition, the nerve cell death inhibitor of the present invention of cut-out inhibitor that contains CaNA is because the prevention or the curative that can be used as with nerve cell death diseases associated such as dementia disease use, and all are very useful.In addition, if use the additive of the cultured cell culture medium of the present invention of the cut-out inhibitor that contains CaNA, also can make cultured cell propagation better.In addition, the cut-out inhibitor of CaNA of the present invention also can be used as the research relevant with nerve cell death and uses with reagent.
Sequence table
<110>Tomizawa，Kazuhito
Matsui，Hideki
＜120〉the persistent activation inhibitor of calcineurin
<130>JP-13650
<160>6
<210>1
<211>16
<212>PRT
＜213〉people
<400>1
Phe?Asp?Gly?Ala?Thr?Ala?Ala?Ala?Arg?Lys?Glu?Val?Ile?Arg?Asn?Lys
1 5 10 15
<210>2
<211>17
<212>PRT
＜213〉people
<400>2
Arg?Glu?Glu?Ser?Glu?Ser?Val?Leu?Thr?Leu?Lys?Gly?Leu?Thr?Pro?Thr
1 5 10 15
Gly
<210>3
<211>521
<212>PRT
＜213〉people
<400>3
Met?Ser?Glu?Pro?Lys?Ala?Ile?Asp?Pro?Lys?Leu?Ser?Thr?Thr?Asp?Arg
1 5 10 15
Val?Val?Lys?Ala?Val?Pro?Phe?Pro?Pro?Ser?His?Arg?Leu?Thr?Ala?Lys
20 25 30
Glu?Val?Phe?Asp?Asn?Asp?Gly?Lys?Pro?Arg?Val?Asp?Ile?Leu?Lys?Ala
35 40 45
His?Leu?Met?Lys?Glu?Gly?Arg?Leu?Glu?Glu?Ser?Val?Ala?Leu?Arg?Ile
50 55 60
Ile?Thr?Glu?Gly?Ala?Ser?Ile?Leu?Arg?Gln?Glu?Lys?Asn?Leu?Leu?Asp
65 70 75 80
Ile?Asp?Ala?Pro?Val?Thr?Val?Cys?Gly?Asp?Ile?His?Gly?Gln?Phe?Phe
85 90 95
Asp?Leu?Met?Lys?Leu?Phe?Glu?Val?Gly?Gly?Ser?Pro?Ala?Asn?Thr?Arg
100 105 110
Tyr?Leu?Phe?Leu?Gly?Asp?Tyr?Val?Asp?Arg?Gly?Tyr?Phe?Ser?Ile?Glu
115 120 125
Cys?Val?Leu?Tyr?Leu?Trp?Ala?Leu?Lys?Ile?Leu?Tyr?Pro?Lys?Thr?Leu
130 135 140
Phe?Leu?Leu?Arg?Gly?Asn?His?Glu?Cys?Arg?His?Leu?Thr?Glu?Tyr?Phe
145 150 155 160
Thr?Phe?Lys?Gln?Glu?Cys?Lys?Ile?Lys?Tyr?Ser?Glu?Arg?Val?Tyr?Asp
165 170 175
Ala?Cys?Met?Asp?Ala?Phe?Asp?Cys?Leu?Pro?Leu?Ala?Ala?Leu?Met?Asn
180 185 190
Gln?Gln?Phe?Leu?Cys?Val?His?Gly?Gly?Leu?Ser?Pro?Glu?Ile?Asn?Thr
195 200 205
Leu?Asp?Asp?Ile?Arg?Lys?Leu?Asp?Arg?Phe?Lys?Glu?Pro?Pro?Ala?Tyr
210 215 220
Gly?Pro?Met?Cys?Asp?Ile?Leu?Trp?Ser?Asp?Pro?Leu?Glu?Asp?Phe?Gly
225 230 235 240
Asn?Glu?Lys?Thr?Gln?Glu?His?Phe?Thr?His?Asn?Thr?Val?Arg?Gly?Cys
245 250 255
Ser?Tyr?Phe?Tyr?Ser?Tyr?Pro?Ala?Val?Cys?Asp?Phe?Leu?Gln?His?Asn
260 265 270
Asn?Leu?Leu?Ser?Ile?Leu?Arg?Ala?His?Glu?Ala?Gln?Asp?Ala?Gly?Tyr
275 280 285
Arg?Met?Tyr?Arg?Lys?Ser?Gln?Thr?Thr?Gly?Phe?Pro?Ser?Leu?Ile?Thr
290 295 300
Ile?Phe?Ser?Ala?Pro?Asn?Tyr?Leu?Asp?Val?Tyr?Asn?Asn?Lys?Ala?Ala
305 310 315 320
Val?Leu?Lys?Tyr?Glu?Asn?Asn?Val?Met?Asn?Ile?Arg?Gln?Phe?Asn?Cys
325 330 335
Ser?Pro?His?Pro?Tyr?Trp?Leu?Pro?Asn?Phe?Met?Asp?Val?Phe?Thr?Trp
340 345 350
Ser?Leu?Pro?Phe?Val?Gly?Glu?Lys?Val?Thr?Glu?Met?Leu?Val?Asn?Val
355 360 365
Leu?Asn?Ile?Cys?Ser?Asp?Asp?Glu?Leu?Gly?Ser?Glu?Glu?Asp?Gly?Phe
370 375 380
Asp?Gly?Ala?Thr?Ala?Ala?Ala?Arg?Lys?Glu?Val?Ile?Arg?Asn?Lys?Ile
385 390 395 400
Arg?Ala?Ile?Gly?Lys?Met?Ala?Arg?Val?Phe?Ser?Val?Leu?Arg?Glu?Glu
405 410 415
Ser?Glu?Ser?Val?Leu?Thr?Leu?Lys?Gly?Leu?Thr?Pro?Thr?Gly?Met?Leu
420 425 430
Pro?Ser?Gly?Val?Leu?Ser?Gly?Gly?Lys?Gln?Thr?Leu?Gln?Ser?Ala?Thr
435 440 445
Val?Glu?Ala?Ile?Glu?Ala?Asp?Glu?Ala?Ile?Lys?Gly?Phe?Ser?Pro?Gln
450 455 460
His?Lys?Ile?Thr?Ser?Phe?Glu?Glu?Ala?Lys?Gly?Leu?Asp?Arg?Ile?Asn
465 470 475 480
Glu?Arg?Met?Pro?Pro?Arg?Arg?Asp?Ala?Met?Pro?Ser?Asp?Ala?Asn?Leu
485 490 495
Asn?Ser?Ile?Asn?Lys?Ala?Leu?Ala?Ser?Glu?Thr?Asn?Gly?Thr?Asp?Ser
500 505 510
Asn?Gly?Ser?Asn?Ser?Ser?Asn?Ile?Gln
515 520
<210>4
<211>11
<212>PRT
＜213〉HIV virus
<400>4
Tyr?Gly?Arg?Lys?Lys?Arg?Arg?Gln?Arg?Arg?Arg
1 5 10
<210>5
<211>26
<212>PRT
＜213〉people
<400>5
Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Phe?Asp?Gly?Ala?Thr?Ala
1 5 10 15
Ala?Ala?Arg?Lys?Glu?Val?Ile?Arg?Asn?Lys
20 25
<210>6
<211>27
<212>PRT
＜213〉people
<400>6
Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Arg?Glu?Glu?Ser?Glu?Ser
1 5 10 15
Val?Leu?Thr?Leu?Lys?Gly?Leu?Thr?Pro?Thr?Gly
20 25
Claims (4)
1. one kind is suppressed the cut-out inhibitor that Calpain cuts off calcineurin A subunit, and it contains the peptide of sequence 1 and/or the peptide shown in the sequence 2.
2. nerve cell death inhibitor is an effective ingredient with the cut-out inhibitor of the described calcineurin A of claim 1 subunit.
A dementia disease carry out inhibitor, be effective ingredient with the cut-out inhibitor of the described calcineurin A of claim 1 subunit.
4. the additive of cell and brain section culture medium contains the cut-out inhibitor of the described calcineurin A of claim 1 subunit.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2002294815 | 2002-10-08 | ||
| JP294815/2002 | 2002-10-08 |
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|---|---|
| CN1691960A CN1691960A (en) | 2005-11-02 |
| CN100340288C true CN100340288C (en) | 2007-10-03 |
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|---|---|---|---|
| CNB2003801005939A Expired - Fee Related CN100340288C (en) | 2002-10-08 | 2003-10-07 | Inhibitors for continuous activation of calcineurin |
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| Country | Link |
|---|---|
| US (1) | US7183375B2 (en) |
| EP (1) | EP1552847A4 (en) |
| JP (1) | JP4642469B2 (en) |
| KR (1) | KR100833728B1 (en) |
| CN (1) | CN100340288C (en) |
| TW (1) | TW200407160A (en) |
| WO (1) | WO2004032955A1 (en) |
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| WO2012145890A1 (en) * | 2011-04-25 | 2012-11-01 | 麦克利科技有限公司 | Use of regulator of calcineurin 1 for manufacturing medicament for treatment of diseases associated with increased nf-κb activity |
| CN117511882A (en) * | 2022-08-03 | 2024-02-06 | 浙江大学医学院附属第一医院 | Universal immune effector cell and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002000872A2 (en) * | 2000-06-29 | 2002-01-03 | Exonhit Therapeutics Sa | Compositions and methods for treating or detecting degenerative diseases of the motor neurons |
-
2003
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- 2003-10-07 CN CNB2003801005939A patent/CN100340288C/en not_active Expired - Fee Related
- 2003-10-07 US US10/518,710 patent/US7183375B2/en not_active Expired - Fee Related
- 2003-10-07 JP JP2004542834A patent/JP4642469B2/en not_active Expired - Fee Related
- 2003-10-07 WO PCT/JP2003/012816 patent/WO2004032955A1/en active Application Filing
- 2003-10-07 EP EP03751353A patent/EP1552847A4/en not_active Withdrawn
- 2003-10-07 KR KR1020057005991A patent/KR100833728B1/en not_active IP Right Cessation
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002000872A2 (en) * | 2000-06-29 | 2002-01-03 | Exonhit Therapeutics Sa | Compositions and methods for treating or detecting degenerative diseases of the motor neurons |
Non-Patent Citations (2)
| Title |
|---|
| activation of a calmodulin-depentdetn phosphatase by aca2+dependetn protease TALLTANT EA et al,Biochemistry,Vol.27 No.6 1988 * |
| Calpain-dependent cleavage of cain/cabin1 activatescalcineurin to mediate calcium-triggered cell death. KIM,M.J. et al,Proc.Natl.Acad.Sci.,Vol.99 No.15 2002 * |
Also Published As
| Publication number | Publication date |
|---|---|
| JP4642469B2 (en) | 2011-03-02 |
| US20060177431A1 (en) | 2006-08-10 |
| KR20050065581A (en) | 2005-06-29 |
| CN1691960A (en) | 2005-11-02 |
| TWI341208B (en) | 2011-05-01 |
| US7183375B2 (en) | 2007-02-27 |
| KR100833728B1 (en) | 2008-05-29 |
| EP1552847A1 (en) | 2005-07-13 |
| TW200407160A (en) | 2004-05-16 |
| EP1552847A4 (en) | 2009-10-28 |
| WO2004032955A1 (en) | 2004-04-22 |
| JPWO2004032955A1 (en) | 2006-02-09 |
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